Effects of tidal fluctuations of salinity on hemolymph composition of the southern oyster drill Thais haemastoma

Effects of varying the amplitude and duration of tidal fluctuations of salinity upon the hemolymph osmotic and ionic composition of the southern oyster drill Thais haemastoma were studied. The composition of diluent was varied during one experiment to approximate Mississippi River water. Snails were also subjected to a diurnal 20-10-20%. S fluctuation pattern for two weeks and hemolymph was collected twice daily. Amplitude of hemolymph osmolality and ion fluctuation during tidal fluctuations of salinity was directly related to the amplitude of ambient salinity fluctuation and inversely related to the rate of fluctuation. The rate of hemolymph osmolality and ion change was directly related to the rate of ambient salinity change. Dilution of seawater with simulated river water instead of deionized water resulted in a reduced amplitude of fluctuation of hemolymph osmolality and ion concentration. Most of the hemolymph osmolality fluctuation was due to solute movement and not to shifts in body water. Hemolymph, sodium and chloride level changed in a similar manner throughout all of the experiments except the 10-5-10% S-simulated river-water experiment in which chloride changed much less than sodium. Hemolymph ninhydrinpositive substance (NPS) levels cycled inversely with ambient salinity during the 30-10-30%. and 20-10-20%. S diurnal and the 30-10-30%. S semidiurnal experiments, but did not change during the 10-5-10%. S deionized water or simulated river-water experiment. Snails fed for most of the 2-week 20-10-2%. S diurnal cycle fluctuation experiment and no mortality occurred. Drills were hyperosmotic to the ambient water at all but two sampling periods, when they were isosmotic. Hemolymph NPS levels tended to be higher during low-salinity slack water than during high-salinity slack water. Even small fluctuations of ambient salinity result in fluctuations of hemolymph osmolality and ionic composition which may affect rate functions within the zone of capacity adaptation of the southern oyster drill.     

Effects of salinity fluctuation on the hemolymph composition of the blue crab Callinectes sapidus

The hemolymph of the blue crab Callinectes sapidus was hyperosmotic during 20-10-20 S and 30-10-30 S diurnal cycles. The hemolymph became isosmotic at 26 S and hyposmotic at 28 S in the 10-30-10 S diurnal cycle. Hemolymph Na⁺ was hyperionic to seawater throughout all cycles. Hemolymph Cl^- was hyperionic below 24 S and either isionic or hypoionic from 24 to 30 S. Hemolymph K⁺ concentrations were hyperionic below 26 S and either isionic or hypoionic from 26 to 30 S. Hemolymph Mg⁺⁺ values were hypoionic over the experimental salinity range (10 to 30). Hemolymph ninhydrin-positive substances (NPS) levels were directly related to ambient salinity.     

Physiological responses of horseshoe crab (<Emphasis Type="Italic">Limulus polyphemus</Emphasis>) embryos to osmotic stress and a possible role for stress proteins (HSPs)

We tested the effects of osmotic stress on survival, developmental rate, and level of HSPs on American horseshoe crab (Limulus polyphemus) embryos. Animals were maintained in the laboratory at an ambient salinity of 20 ppt and then exposed to 4-h osmotic shocks at salinities of 10, 30, 40, 50, and 60 ppt, with a control group at 20 ppt. Horseshoe crab embryos had 100% developmental success (defined as individuals reaching the first instar or trilobite larval stage) at all salinities. However, osmotic stresses, especially hyperosmotic conditions, slowed the rate of development. Embryos subjected to osmotic stress showed higher levels of HSP70 and HSP90 than control animals kept at a salinity of 20 ppt. HSPs are of value to horseshoe crab embryos in surviving the fluctuating salinities that are typical of estuarine beach habitats.     

Osmotic and chloride regulation in the hemolymph of the tiger prawn Penaeus monodon during molting in various salinities

The effect of molting on osmotic and chloride concentrations in the blood of the prawn Penaeus monodon Fabricius (20±3 g) at various salinities was investigated. Prawns were obtained from ponds in Iloilo, Philippines, in 1984. They were stocked in salinities of 8, 20, 32 and 44, and their hemolymph was sampled during molt (Time 0), and then 0.125, 0.25, 0.5, 1, 2, 4, 6, 10 and 14 d after molting. Prawns during and immediately after molt tended to conform to the environmental osmolality. Subsequent postmolt (0.5 d) stages displayed more divergence from external salinity. The isosmotic point was higher (940±30 mOsm kg^-1) during molt than during intermolt (663±8 mOsm/kg^-1), suggesting different osmotic requirements in early molt. Hyperregulation of hemolymph chloride below 20 S, as well as isoionic point (301±6 mM), were independent of molting stage. At 20 S and above, newly molted (0 to 0.25 d post-molt) individuals tended to conform to the external chloride concentration while intermolt (0.5 d) post-molt individuals did not. Contribution of hemolymph chloride to hemolymph osmolality was greater during intermolt than during ecdysis, suggesting an important role for other negatively charged ions during molt. When molt occurred in 20 S (the test salinity most similar to the isoionic salinity), there was little or no change in hemolymph osmolality or chloride concentration from 0 to 14 d postmolt. At 8, 32 and 44 S, the change from molt to intermolt values in hemolymph osmotic and chloride concentrations was hyperbolic. Non-linear least-squares regression showed that prawns generally achieved intermolt values within 1 d after molting. Prawns at intermolt regulated hemolymph osmolality (620 to 820 mOsm kg^-1) and chloride concentration (300 to 450 mM) at a much narrower range than during molt (520 to 1 170 mOsm kg^-1 and 250 to 520 mM, respectively). Hemolymph osmolality was a more sensitive indicator of physiological response than hemolymph chloride concentration. Distribution and culture of P. monodon might be limited in low salinities by its ability to maintain a hemolymph osmolality 500 mOsm kg^-1 during molt and 600 mOsm kg^-1 in intermolt, and in high salinities by its capacity to reduce the hemolymph osmolality from values at molt to those in intermolt. Osmotic and chloride concentrations in the blood of P. monodon clearly varied with both molt stage and salinity of the medium. Dependence on external factors, however, gradually declined in older molt stages, suggesting a reduction in integument permeability and greater development of ion absorption/secretion mechanisms as the exoskeleton hardened.SEAFDEC Contribution No. 197     

Osmoregulation of the grass shrimp Palaemonetes pugio exposed to polychlorinated biphenyls (PCBs). I. Effecton chloride and osmotic concentrations and chloride-and water-exchange kinetics

Grass shrimp, Palaemonetes pugio, were capable of hypo- and hyper-osmotic regulation of body fluids. Hemolymph chloride and osmotic concentrations were maintained at relatively stable levels over a wide salinity range. Following an abrupt transfer from intermediate (14 and 17) to high (31 and 35) or low (1 and 2) salinities, hemolymph chloride levels exhibited initial overshoot and undershoot, respectively, of new steady-state levels. Osmotic concentrations exhibited an initial undershoot at low, but not overshoot at high salinity. Chloride space in salinity-acclimated shrimp was relatively stable at salinities from 1 to 35. Changes in chloride space following salinity transfer paralleled those of hemolymph chloride levels, and are discussed in the light of alterations in intracellular sodium concentrations reported earlier. Rate constants for chloride turnover indicated independent exchanges of sodium and chloride ions. Water-turnover measurements showed that permeability of P. pugio was greatest at the isosmotic salinity (17) and reduced at salinities which were associated with active osmoregulation. Exposure to sublethal and 96-h LC₅₀ levels of Aroclor^® 1254 did not seriously alter hemolymph chloride and osmotic concentrations, chloride space or chloride-exchange kinetics in adult shrimp. Disruption of hemolymph chloride regulation in juvenile shrimp was associated with large mortalities not observed in adults. Shrimp exposed to Aroclor 1254 at 17 S exhibited reduced water permeability similar to levels previously observed in controls at high and low salinities in response to osmotic or ionic gradients. Exposure to PCBs did not result in further reduction in permeability at the latter salinities.     

The effect of organic material and inorganic ions on the photosynthetic rate of the red alga Bostrychia binderi from a Florida estuary

Various constituents of spring water (calcium, bicarbonate, nitrate, phosphate, total organic material) influence the response of photosynthetic rate of Bostrychia binderi Harvey to changes in salinity. The rate of photosynthesis increased with a decrease in salinity. The rate of photosynthesis in low salinities was greater in seawater diluted with spring water than in sewater diluted with distilled water. Elevation of photosynthetic rates in the lower salinities (0 and 5 ppt) was partially due to increased levels of bicarbonate and various nutrients present in natural spring water. The higher calcium levels in spring water resulted in higher photosynthetic rates in plants held for 3 to 7 d in the lower salinities (0 to 5 ppt). Increased levels of calcium in salinities of 5 ppt or higher increased the photosynthetic rate only during the first 7 d of exposure, since acclimation occurred equally in individuals held for 2 to 8 wk in sewater diluted with distilled or spring water. This study suggests that the diverse algal floras, characteristic of estuaries on the west coast of Florida are in part the result of natural spring water mixing with seawater, sustaining the algae over short periods of low salinities.     

Egg mortality and hatching rate of Baltic cod (Gadus morhua) in different salinities

Egg mortality of Baltic cod (Gadus morhua L.), collected off northern Gotland, Sweden, in 1990, was studied in four different salinities — 10 and 15 ppt (salinity of the principal spawning areas of Baltic cod) and 5 and 7 ppt (salinity above the halocline) — in laboratory experiments. Mortality was high during the first 4 d of development, but after gastrulation mortality was low in all salinities tested, except for 5 ppt, in which mortality increased slightly before hatching. Mortality during hatching varied considerably with salinity. No hatching occurred in 5 ppt salinity, and only a few larvae survived in 7 ppt salinity; in contrast, mortality during hatching was comparatively low in salinities of 10 and 15 ppt.     

Suitability of estuarine nursery zones for juvenile weakfish (Cynoscion regalis): effects of temperature and salinity on feeding,growth and survival

Juvenile weakfish, Cynoscion regalis (Bloch and Schneider, 1801), exhibit significant spatial diffrences in growth rate and condition factor among estuarine nursery zones in Delaware Bay. The potential influence of temperature and salinity on the suitability of estuarine nursery areas for juvenile weakfish was investigated in laboratory experiments by measuring ad libitum feeding rate, growth rate and gross growth efficiency of juveniles collected in Delaware Bay in 1990 (40 to 50 mm standard length; 1.4 to 2.1 g) in 12 temperature/salinity treatments (temperatures: 20, 24, 28°C; salinities: 5, 12, 19, 26 ppt) representing conditions encountered in different estuarine zones during spring/summer. Feeding rates (FR) increased significantly with temperature at all salinities, ranging from 10 to 15% body wt d^-1 at 20°C to 33–39% body wt d^-1 at 28°C. Specific growth rates (SGR) ranged from 1.4 to 9.4% body wt d^-1 (0.3 to 1.5 mm d^-1) and gross growth efficiencies (K ₁) varied from 13.6 to 26.4% across temperature/salinity combinations. Based on nonlinear multiple regression models, predicted optimal temperatures for SGR and K ₁ were 29 and 27°C, respectively. Salinity effects on SGR and K ₁ were significant at 24 and 28°C where predicted optimal salinity was 20 ppt. At these warmer temperatures, SGR and K ₁ were significantly lower at 5 than at 19 ppt despite higher FR at 5 ppt. Therefore, maximum growth rate and growth efficiency occurred under conditions characteristic of mesohaline nurseries. This finding is consistent with spatial patterns of growth in Delaware Bay, implying that physicochemical gradients influence the value of particular estuarine zones as nurseries for juvenile weakfish by affecting the energetics of feeding and growth. Laboratory results indicate a seasonal shift in the location of physiologically optimal nurseries within estuaries. During late spring/early summer, warmer temperatures in oligohaline areas permit higher feeding rate and faster growth compared to mesohaline areas. By mid-late summer, spatial temperature gradients diminish and mesohaline areas provide more suitable physicochemical conditions for growth rate and growth efficiency whereas oligohaline areas become energetically stressful. Substantial mortality occurred at 5 ppt and 28°C, providing additional evidence that oligohaline conditions are stressful during late summer. Furthermore, juveniles provided a choice among salinities in laboratory trials preferred those salinities which promoted higher growth rates. The extensive use of oligohaline nurseries by juvenile weakfish despite the potential for reduced growth rate and growth efficiency suggests this estuarine zone may provide a substantial refuge from predation.     

Active salinity choice and enhanced swimming endurance in 0 to 8-d-old larvae of diadromous gobies,including Sicydium punctatum (Pisces), in Dominica,West Indies

We studied the early life history of diadromous gobies in Dominica, West Indies, from May 1989 to May 1991, emphasising Sicydium punctatum Perugia. The transition of newly hatched larvae from upriver nest sites to the sea was studied in laboratory experiments. Newly hatched larvae are negatively buoyant but avoid settling to the bottom by active swimming during drift to the sea. Laboratory experiments evaluated salinity preferences and effects on swimming endurance. Larvae in haloclines actively selected low to intermediate salinities. Initially (0 to 5-d post-hatch), larvae minimized exposure to salinities >10 ppt, but later (5 to 8-d) occupied increasingly saline water. Larvae in no-choice freshwater or seawater treatments ceased activity at 4 to 5 d, but in haloclines larvae remained active up to 8 d post-hatch. Salinities <10 ppt are important for early survival of sicydiine gobies. Implications for larval survival and transport are discussed.     

Effect of salinity adaptation on nitrogen metabolism in the freshwater fish Tilapia mossambica. I. Tissue protein and amino acid levels

Water, proteins and total free amino acids were estimated in different tissues of the euryhaline fish Tilapia mossambica after adaptation to various strenghts of sea water. The water content did not vary significantly in any tissue on salinity adaptation. The soluble and insoluble proteins displayed a general and considerable decrease in muscle, liver and heart; the decrease in the soluble fraction in the heart and the proteins of the muscle in 75% sea water (100% sea water=32.5 S) were significant. The gill proteins did not alter with salinity; the kidney proteins tended to increase slightly in 100% sea water (SW). The total free amino-acid content decreased insignificantly in all tissues on adaptation to 25% SW; in higher salinities, however, the content increased significantly. This increase was sudden and steep in 50% SW, and gradual and less steep in 75 and 100% SW. It is suggested that constancy in water content may contribute to the great adaptability of T. mossambica to heterosmotic media, and that the total free amino acids may be involved in isosmotic intracellular regulation. The possibility of amino acid increase as a result of protein breakdown is also indicated.     

The effect of pulsed versus gradual salinity reduction on the physiology and survival of Halophila johnsonii Eiseman

Plants of Halophila johnsonii Eiseman were exposed, in mesocosms, to either pulsed hyposalinity treatments of 30, 15, 10, and 8 or gradual salinity reductions of two every 2?days. When salinity was pulsed, survivorship (>80?%) and maximum quantum yields (>0.7) were high in the 30 and 15 salinity treatments, but both declined in the 10 and 8 salinity treatments. Leaf osmolality declined with respect to salinity treatment, but the difference between leaf and media osmolality remained relatively constant (675?±?177?mmol?kg^?1). In contrast, when salinity was gradually reduced, survivorship remained high from salinities of 30 to 4, and maximum quantum yields remained high from salinities of 30 to 6. Leaf osmolality declined linearly with respect to media osmolality and, similar to the pulsed treatments, the difference between leaf and media osmolality remained relatively constant from salinities of 30 to 2 (638?±?161?mmol?kg^?1). Trolox equivalent antioxidant capacity declined over time in both pulsed and gradual salinity reduction. The results indicate that H. johnsonii is more tolerant of hyposalinity than has previously been reported and that gradually reducing salinity extended its low-salinity tolerance threshold by approximately a salinity of 10.     

Effects of salinity and adult extract on settlement of the oligohaline barnacle Balanus subalbidus

Balanus subalbidus (Henry) has the most oligohaline distribution of three congeneric barnacles in Chesapeake Bay and tolerates prolonged exposure to fresh water. We studied larval settlement (i.e., permanent attachment and metamorphosis) of B. subaldius in the laboratory, over a 3 yr period, May 1989 to March 1992, under the following conditions: (1) across an array of salinities at 25°C in the presence and absence of settlement factor consisting of adult B. subalbidus extract; (2) in the presence of conspecific or congeneric settlement factors; and (3) cyprids which were, and were not, induced to delay metamorphosis were compared in their capacities to settle in a range of salinities. Discrepancies between salinity profiles of larval settlement in the laboratory and adult oligohaline distribution in the estuary were striking, and there was a significant interaction between salinity and settlement factor. Averaging results of four different batches of larvae, although peak settlement (87±9%) of B. subalbidus occurred at 2 ppt salinity in the presence of adult cue, substantial settlement also occurred at higher salinities: >70% at 5, 10 and 15 ppt; and 47% at 20 and 25 ppt. In addition, settlement in the absence of settlement factor was relatively high (>50%) and peaked at mid-salinity ranges (e.g. 56±10% at 15 ppt). Variation observed in settlement among larval batches reflected detailed differences in settlement between adjacent test salinities. No difference in settlement occurred between replicate aliquots of cyprids within a batch. Cyprids of B. subalbidus settled most abundantly in the presence of settlement factor extracted from conspecifics, followed in decreasing order by settlement factor extracted from B. improvisus and B. eburneus. Delay of metamorphosis produced by keeping B. subalbidus cyprids for 8 d at 5°C resulted in a decreased level of settlement, but settlement frequency patterns of delayed and non-delayed cyprids were indistinguishable relative to salinity. These results indicate that the oligohaline distribution of adult B. subalbidus is probably not determined by larval behavior at settlement. We suggest that pre-settlement behavior, resulting in larval retention in low saline waters, could be an important factor in determining distribution of this species.     

Water balance systems of Rangia cuneata: ionic and amino acid regulation in changing salinities

In order to study the interaction of the extracellular and intracellular osmoregulatory systems of the bivalve Rangia cuneata, we have measured blood osmotic and ionic concentrations together with intracellular free amino acid concentrations and total tissue water under identical salinity conditions. Like freshwater bivalves, the blood of R. cuneata is maintained hyperosmotic (50 mOsm) to the environment in salinities below 110 mosm by the regulation of Na⁺, Cl^-, K⁺ and Ca²⁺ concentrations. On the other hand in company with marine bivalves, R. cuneata also regulates intracellular free amino acids (FAA) as a mechanism to control cellular volume during osmotic stress over the entire non-lethal salinity range (3 to 620 mOsm). Alanine is the predominant intracellular osmotic effector. Thus, by utilizing the osmoregulatory mechanisms of both marine and freshwater bivalves, R. cuneata is able to tolerate salinities ranging from freshwater to 25 ppt and to traverse the faunal salinity boundary, known as the horohalinicum (5 to 8 ppt), controlling cell volume throughout.Please address requests for reprints to Dr. S. K. Pierce     

Effect of environmental factors on byssal thread formation

The effects of various factors on byssal thread formation have been examined in the laboratory using Modiolus demissus and Mytilus edulis. With M. demissus thread formation (threads/mussel/h0 and proportion of mussels forming threads decreased with increasing size; prior exposure to air enhanced subsequent thread formation; mechanical agitation reduced thread formation. In addition, low salinity acclimated mussels adapted more rapidly to 32 ppt than high salinity acclimated mussels did to 16 ppt; threads were not formed in the absence of calcium and/or magnesium; and there was no reduction in thread formation at temperatures as high as 27° to 28°C. M. edulis died and thread formation approached zero at temperatures exceeding 26°C.     

Effects of sudden changes in salinity on endogenous rhythms of the spotted sea bass Lateolabrax sp.

The endogenous rhythm of oxygen consumption in juvenile spotted sea bass (Lateolabrax sp.) was measured to test the effects of sudden changes in salinity on the metabolic activity. Mean oxygen consumption rates of this euryhaline fish decreased by 13.5 to 16.0% and 25.3 to 36.4% when they were transferred from 31.5 to 15‰ seawater and to fresh water (0‰), respectively. The maximum rate of oxygen consumption was observed between 18:00 and 19:00 hrs local time, 1 to 2 h before sunset, even though they were kept in constant darkness. The peaks of oxygen consumption occurred in 23.2- and 23.3-h intervals, which correspond with a circadian rhythm, as revealed by maximum entropy spectral analysis. A markedly weakened rhythm in oxygen consumption occurred from 8 to 10 d after onset of the experiments. This study indicates that spotted sea bass can withstand sudden drops in salinity from 31.5‰ to fresh water, and yet maintain a regular though somewhat dampened endogenous rhythm of oxygen consumption. Received: 16 June 1997 / Accepted: 3 February 1998     

A comparison of transport-related gill enzyme activities and tissue-specific free amino acid concentrations of Baltic Sea (brackish water) and freshwater threespine sticklebacks, Gasterosteus aculeatus, after salinity and temperature acclimation

The osmoregulatory abilities of one freshwater and two brackish water (Baltic Sea) populations of the euryhaline teleost fish Gasterosteus aculeatus were studied with respect to evolutionary physiology. Plasma osmolality, activities of Na⁺K⁺-ATPase, citrate synthase, creatine kinase in the gill and free amino acids in liver, axial muscle and pectoral fin muscle were measured. After transfer from 10 to 35 ppt at 15 °C, time-course changes of plasma osmolality and gill Na⁺K⁺-ATPase showed no significant fundamental differences between the freshwater and one of the Baltic Sea populations. In a multi-factorial experiment, each population was exposed to four different abiotic regimes. Both brackish water populations had high mortality in freshwater at 4 °C, which is discussed as a failure of osmotic regulation (reduced taurine concentrations). Freshwater specimens had higher levels of glycine in the axial and pectoral fin muscles compared to the brackish water populations. This is interpreted as a genetically based effect. In brackish (20 ppt) water of 15 °C, the freshwater population had high activities of Na⁺K⁺-ATPase, but low activities of creatine kinase, whereas both brackish water populations behaved in the opposite way. A fundamental difference between the freshwater and brackish water populations on the level of the osmoregulatory machinery was not observed. Received: 10 December 1998 / Accepted: 22 September 1999     

Feeding ecology of the grooved tiger shrimp <Emphasis Type="Italic">Penaeus semisulcatus</Emphasis> De Haan (Decapoda: Penaeidae) in inshore waters of Qatar,Arabian Gulf

The feeding ecology of the green tiger shrimp Penaeus semisulcatus was studied in inshore fishing grounds off Doha, Qatar, using a combination of stable isotope (δ¹³C and δ¹⁵N) analysis and gut contents examination. Samples of post-larvae, juvenile and adult shrimp and other organisms were collected from intertidal and subtidal zones during the spawning season (January–June). Shrimp collected from shallow water seagrass beds were mostly post-larvae and juveniles and were significantly smaller than the older juveniles and adults caught in deeper macroalgal beds. Gut content examination indicated that post-larvae and juvenile shrimp in seagrass beds fed mainly on benthos such as Foraminifera, polychaetes, benthic diatoms and small benthic crustaceans (amphipods, isopods and ostracoda), whereas larger shrimp in the macroalgal beds fed mainly on bivalve molluscs and to a lesser extent polychaetes. In shrimp from both seagrass and algal beds, unidentifiable detritus was also present in the gut (18, 32%). δ¹³C values for shrimp muscle tissue ranged from −9.5 ± 0.26 to −12.7 ± 0.05‰, and δ¹⁵N values increased with increasing shrimp size, ranging from 4.1 ± 0.03 to 7.7 ± 0.11‰. Both δ¹⁵N values and δ¹³C values for shrimp tissue were consistent with the dietary sources indicated by gut contents and the δ¹³C and δ¹⁵N values for primary producers and prey species. The combination of gut content and stable isotope data demonstrates that seagrass beds are important habitats for post-larvae and juvenile P. semisulcatus, while the transition to deeper water habitats in older shrimp involves a change in diet and source of carbon and nitrogen that is reflected in shrimp tissue stable isotope ratios. The results of the study confirm the linkage between sensitive shallow water habitats and the key life stages of an important commercially-exploited species and indicate the need for suitable assessment of the potential indirect impacts of coastal developments involving dredging and land reclamation.     

Indicators of physiological and immunological status of <Emphasis Type="Italic">Litopenaeus setiferus</Emphasis> wild populations (Crustacea,Penaeidae)

Constant environmental changes in oceanic and estuarine systems due to human activity in coastal zones require different tools and strategies for their study to be able to assess the health status of aquatic ecosystems. The Gulf of Mexico has a tremendous ecological importance because of its biological diversity. Baseline levels of blood metabolites and immune components were defined for wild tropical populations of Litopenaeus setiferus, an important shrimp species of the Gulf of Mexico. Osmotic pressure, oxy hemocyanin (OxyHc), glucose, proteins, cholesterol, lactate, and triacylglycerols (TAG) were used as indicators of physiological status; pro phenoloxydase (proPO) and phenoloxydase (PO) quantification of hemocytes, as well as respiratory burst, were used to assess the immunological status. Significant differences were observed in live weight of juveniles, males, and females of L. setiferus with mean values of 10.54, 35.05, and 40.33 g, respectively. A value of 460 mOsm/kg was recorded in the water where juveniles were sampled and 1,000 mOsm/kg in adult sampling zones. Consequently, the osmotic pressure of L. setiferus juveniles (718.02 mOsm/kg) was 22% lower than in adults (925 mOsm/kg); there were no differences between sexes. A significantly lower value in juvenile OxyHc was observed. This value was 24 and 16% lower than that observed in males and females, respectively. Hemolymph glucose, cholesterol, and triacylglycerol levels were not statistically different between shrimp stages, showing median values of 0.19, 0.32, and 0.39 mg/ml of glucose, cholesterol, and triacylglycerols, respectively. Hemocyanin was distributed normally in the juvenile–male population but not in females. A proportion between 52 to 81% of OxyHc of the total protein (OxyHc/Prot) was obtained for all stages. Digestive gland glycogen showed no normal distribution in juveniles and adults. Digestive gland glycogen was significantly higher in juveniles (median value of 2.64 mg/g) than in adults (median value of 1.58 mg/g). A higher value of lactate (0.11 mg/ml) was observed in juveniles than in males (0.09 mg/ml) and females (0.04 mg/ml). Granular cells (GC) corresponded to 36, 44, and 39% of the total hemocytes (TH) observed in juveniles, males, and females. ProPO showed no normal distribution in all stages. A high proPO activity was recorded in L. setiferus females that was 95% higher than in the juvenile–male group. Respiratory burst was divided into two groups, juveniles–males and females, and was distributed normally in both. Female respiratory burst values were 40 and 45% higher than those observed in the juvenile and male groups, respectively. There was an increment in OxyHc, proteins, PO, hemocytes, and respiratory burst with an increase in body weight. In contrast, the relationship of live weight to glucose, TAG, and proPO revealed no significant regressions. A significant decrease in cholesterol, glycogen, and lactate with an increase in body weight was also observed. We now have indicators of nutritional and immune status of juveniles and adults that can reveal changes in trophic relations and health at the population level. This will serve as a basis for using L. setiferus as environmental sentinels, as non-controversial and inexpensive models to evaluate the potential changes that human activities could exert on this important marine ecosystem.Communicated by P.W. Sammarco, Chauvin     

Effects of tidal fluctuations of salinity on pericardial fluid composition of the American oyster Crassostrea virginica

Crassostrea virginica Gmelin were subjected to simulated tidal fluctuations of salinity, and the subsequent effects on osmotic and ionic composition of the pericardial fluid, body water and valve movements were investigated. Ambient salinity fluctuation patterns of 20-10-20, 15-10-15 and 10-5-10 were simulated during 24.8-h periods. An additional 10-5-10 S experiment was performed using a dilution water approximating the ionic composition of Mississippi River water with regard to Mg⁺⁺, Ca⁺⁺ and SO ₄ ⁼ , instead of deionized water. Finally the effects of a 2-week diurnal fluctuation pattern between 20 and 10 S were investigated with respect to pericardial fluid composition. Pericardial fluid osmolality, concentrations of Cl^-, Na⁺, Mg⁺⁺, K⁺, Ca⁺⁺ and ninhydrin-positive substances (NPS) were analyzed periodically throughout all experiments. Pericardial fluid osmolality was slightly hyperosmotic as ambient water salinity decreased during a cycle, and then became slightly hyposmotic as ambient salinity increased. In the 2-week experiment, pericardial fluid osmolality tracked ambient seawater closely through Day 5, but became more intermediate between high and low seawater values as the experiment progressed. Similar patterns during fluctuations of salinity were observed for Na⁺, Cl^-, Mg⁺⁺ and Ca⁺⁺. Pericardial fluid K⁺ levels did not track ambient seawater as closely as did other ions. The ionic composition of dilution water had little effect on the osmotic or ionic response of the oyster's pericardial fluid. Pericardial fluid NPS level varied inversely with salinity during the 20-10-20 cycle. During the longterm fluctuation experiment, NPS values gradually decreased over the 2-week period compared to constant salinity control values. Percent body water also varied inversely with ambient salinity. Solute movement accounted for most of the change in pericardial fluid osmolality during the simulated cycles with water movement responsible for 1 to 11%. Water movement contributed more to the change of pericardial fluid osmolality during the decreasing salinity phase than the increasing phase of a given cycle. During 20-10-20 S cycles, oyster valves remained open 56% of the time (n=23). In contrast, when salinity was abruptly changed from 20 to 10 within 5 min, valve closure occurred in 4.8±0.3 min (n=20). Valves did not reopen for 19.3±1.2 h (n=15).     

Different salinity tolerance mechanisms in Atlantic and Chesapeake Bay conspecific oysters: glycine betaine and amino acid pool variations

Crassostrea virginica (Gmelin) collected in 1989 from several sites within the Chesapeake Bay have narrower salinity tolerances than conspecific oysters collected in 1989 from several Atlantic coast sites (Georgia to Cape Cod). The basis of this physiological difference appears to be the biochemical mechanisms that control cellular osmolality following salinity stress. When adapted to the same salinity, the amino acid pools of both gill and adductor muscles of Atlantic oysters are larger than those of Bay oysters and different in composition. The Atlantic oyster tissues rely primarily on taurine for salinity tolerance, while the Bay oyster tissues have relatively less taurine, depending instead upon alanine, glycine and proline to adapt to high salinity. In addition, Atlantic oyster gill and adductor have 10 to 25 times the glycine betaine concentrations of these tissues from Bay oysters, depending upor the salinity of acclimation. The betaine concentration varies with salinity in Atlantic oysters, but does not change in Bay oysters. The results suggest that these biochemical differences are the basis of the narrower salinity tolerance in Bay oysters. The biochemical differences may reflect genetic differences between Bay and Atlantic oysters.